上海复旦大学附属儿科医院急性下呼吸道感染患儿常见病毒的检测及临床研究

目的 建立套式反转录-聚合酶链反应(nested-RT-PCR)检测临床样本鼻病毒(human rhinovirus,HRV)基因的方法和直接免疫荧光(DFA)的方法对常见呼吸道病毒病原进行检测,了解上海儿科医院小儿急性下呼吸道感染(ALRTI)中病毒感染的现状.方法 收集住院确诊为ALRTI患儿的深部鼻咽分泌物(NPS)标本.用套式RT-PCR的方法检测NPS中的HRV基因,并进一步对扩增产物进行序列分析;用DFA的方法检测NPS中的呼吸道合胞病毒(RSV)、流感病毒(IFV)、副流感病毒(PIV)、腺病毒(ADV)抗原.并分析各种病毒所致ALRTI的临床特点及流行特征.结果 342例患儿除细菌感染外共130例病毒抗原阳性,其中检测出RSV 64例(18.70%),1岁以下婴儿的RSV检出数为71.8%.RSV 8月份起检出率逐渐升高,到12月份达到最高;HRV阳性患儿46例(13.45%),3岁以下婴幼儿检出例数有38例,占HRV检出数的82.6%.HRV致ALRTI全年可见,在3到5月为最高峰;ADV感染9例(2.63%);PIV感染8例(2.3%);IFV感染7例(2.0%);病毒合并感染共有4例.46例HRV阳性标本中选取15份用另1对引物扩增并测序,测得的核苷酸序列经过blast比较,与GenBank中已知HRV序列的同源性在83%～97%,样本间的序列比较提示变异性较大.130例病毒性ALRTI患儿绝大部分诊断为支气管肺炎.以中等度发热为主,末梢血白细胞＜10×109/L 93例(71.5%),CRP＜8 mg/L 99例(76.2%),符合病毒性肺炎的特点,并发症较少,均治愈出院.结论 本地区小儿病毒性ALRTI中,以RSV及HRV占为主要地位,ADV、PIV及IFV相对少见.较少有混合性病毒感染.HRV所致小儿ALRTI以婴幼儿居多,集中于春、秋两季.套式RT-PCR检测HRV基因快速、操作简便、特异性较强,HRV基因组具有高度变异的特点.     

2011年甘肃地区急性呼吸道感染儿童患者中鼻病毒感染规律的研究

探讨人鼻病毒(Human rhinovirus,HRV)在甘肃地区急性呼吸道感染儿童患者中的流行特征。2011年1月至12月,从甘肃地区急性呼吸道感染儿童患者中采集咽拭子标本286份,用多重PCR方法对常见呼吸道病毒性病原体进行筛查。多重PCR结果中HRV阳性的标本再用nested-PCR分别扩增HRV的VP1和VP4/VP2结合区基因片段,阳性结果进行序列测定和基因分析。286份咽拭子标本中,27份标本多重PCR检测结果显示为HRV阳性,检出率为9.44%。本研究中,HRV感染病例的平均年龄为3岁,其中,1岁以内儿童感染比例最高,占总阳性数的44.4%(12/27);在27例HRV阳性病例中,男性占16例(16/185,8.64%),女性占11例(11/101,10.89%),两者无显著性差异;HRV感染的高峰出现在5月份(6/27,22.2%);从HRV感染的疾病构成看,普通感冒的比例最高,占12.24%(12/98);其次为肺炎,占8.50%(13/153);剩余2病例均为支气管炎。基因序列特征分析发现:A组HRV是甘肃地区2011年的优势流行基因型,占阳性病例的84.62%(22/26),其次为C组HRV,占11.54%(3/26),B组HRV仅发现1例。HRV感染在2011年甘肃地区全年均可发生,主要感染以1岁以内儿童。A组HRV是该地区2011年急性呼吸道感染儿童患者中的主要流行基因型,并存在多个基因型和多个HRV血清型共同流行的特点。     

苏浙闽地区婴幼儿急性腹泻患者病毒检测及 临床特征分析

目的分析苏浙闽地区婴幼儿急性腹泻患者常见病毒及患儿临床特征,为婴幼儿急性腹泻的流行病学研究和临床治疗提供参考。方法选取2010至2014年苏浙闽地区多家医院门诊就诊的急性腹泻患儿1 452例,采用ELISA法检测A组轮状病毒(HRV),多重PCR法检测诺如病毒(NoV)、肠道腺病毒(EAdV)和星状病毒(HAstV),并用RT-PCR法对A组轮状病毒阳性的样本进行G和P分型。结果 1 452例患儿中由常见病毒引起的急性腹泻患儿有707例,占总调查患者的48.69%。急性腹泻患儿常见病毒感染中单一病毒感染占90.10%(637/707),其感染病毒的构成以轮状病毒最多(47.52%,336/707),其次为诺如病毒(36.07%,255/707)以及星状病毒(2.40%,17/707)和肠道腺病毒(4.10%,29/707)。病毒感染腹泻患儿中多重感染者共70例,占9.90%(70/707),以A组轮状病毒合并诺如病毒感染最为常见,占多重感染者的68.57%(48/70)。226份A组轮状病毒阳性标本中,G分型以G1为优势分型,占33.19%;P分型中则以P [11]比例最高,占38.50%;G、P型组合以P [11]G11为主(38.50%)。结论 A组轮状病毒是引发婴幼儿急性腹泻的主要病毒,诺如病毒次之,且婴幼儿病毒性腹泻患者可合并多种病毒感染。不同地区腹泻患儿病毒感染阳性率有差异,轮状病毒基因型表现则具有多样性。     

神经内科住院患者医院感染的危险因素及防护措施

目的:了解神经内科住院患者医院感染发生状况,分析致感染的危险因素,为探讨预防控制措施提供参考依据。方法:回顾性分析神经内科3707例患者临床资料,分析确诊为感染患者的感染率、感染部位、病原菌分布,以及医院感染的危险因素,采用SPSS19.0软件对数据进行统计分析。结果:3707例患者发生医院感染193例,感染率为5.21%;感染患者以下呼吸道感染为主,占55.56%,其次为泌尿道感染占21.74%、上呼吸道感染占12.56%;年龄≥65岁、男性、合并基础疾病、接受侵入性操作、一季度、四季度、无意识状态、糖皮质激素使用、广谱抗菌药物使用及住院时间≥21 d的患者医院感染发生率明显高,是引起医院感染的主要危险因素;共检出致病菌211株,其中多重耐药菌126株占59.72%;革兰阴性杆菌141株占66.82%,革兰阳性菌50株占23.70%,真菌20株占9.48%。结论:神经内科患者医院感染发病率较高,导致医院感染的危险因素多,临床中应综合预防控制,提高整体预防控制效果。     

婴儿巨细胞病毒感染31例临床及治疗观察

目的：探讨婴儿巨细胞病毒(CMV)感染的临床特点及治疗。方法：对经酶免疫斑点技术确诊的31例小儿CMV感染,总结其临床特点并选用丙氧鸟苷治疗。结果：高胆红素血症占58．06％,支气管肺炎占25．80％,中枢神经系统损害占16．13％。31例均给丙氧鸟苷治疗,疗程3～8w,结果25例痊愈,5例好转,1例死亡。结论：婴儿巨细胞病毒感染以高胆红素血症、支气管肺炎及中枢神经系统损害为主;丙氧鸟苷是治疗小儿CMV感染的首选药物。     

城关社区成人水痘并发症76例诊治分析

目的了解成人水痘并发症的临床特点,为成人水痘的预防和临床诊治提供依据。方法回顾性分析2011~2015年我院收治的76例成人水痘并发症患者的临床资料,对临床流行特征和并发症发生情况及预后进行分析。结果 76例成人水痘并发症患者继发皮肤感染占84.21%,肝功能损害占55.26%,呼吸道感染占47.37%;发病比例男性高于女性,且以冬春季节发病为主。76例患者经临床应用阿昔洛韦联合麻疹减毒活疫苗治疗,均治愈。结论成人水痘发病率有增高趋势,而且成人水痘并发症有发热程度和热程、全身毒血症状以及皮疹数量、各种并发症发生比例均较儿童高且严重,无疫苗免疫史患者病情较重。临床上应尽早明确诊断,及时治疗;可通过接种水痘减毒活疫苗(VarV)(建议推行2剂次接种的免疫策略)来控制发病率和减少并发症发生。     

呼吸道感染患者多重耐药菌肺炎克雷伯菌的耐药及危险因素分析

摘要 目的：探讨与分析呼吸道感染患者多重耐药菌肺炎克雷伯菌的耐药及危险因素。方法：选择2015年1月到2020年2月本院诊治的呼吸道感染患者65例作为研究对象,收集患者的临床样本进行细菌分离与耐药分析,调查患者的临床资料并进行危险因素分析。结果：在呼吸道感染患者65例中,分离出多重耐药菌肺炎克雷伯菌32株,占比49.2 %,其中下呼吸道、上呼吸道、灌洗液、血液标本分别占50.0 %、9.4 %、25.0 %、6.3 %。32株多重耐药菌肺炎克雷伯菌对头孢曲松、头孢呋辛、氨苄西林、头孢吡肟、头孢噻肟的耐药率分别为71.9 %、87.5 %、96.9 %、84.4 %、81.3 %,对阿米卡星、头孢替坦、左氧氟沙星、亚胺培南、环丙沙星的敏感率分别为59.4 %、68.8 %、81.3 %、75.0 %、81.3 %。非条件 Logistic回归分析显示血型A型、碳青霉烯类抗菌药物使用、引流、机械通气、糖尿病等为导致多重耐药菌肺炎克雷伯菌感染的独立危险因素(P<0.05)。结论：多重耐药菌肺炎克雷伯菌感染在呼吸道感染患者中比较常见,对头孢呋辛、氨苄西林的耐药率比较高,对左氧氟沙星、环丙沙星的敏感率比较高,血型A型、碳青霉烯类抗菌药物使用、引流、机械通气、糖尿病等为导致多重耐药菌肺炎克雷伯菌感染的独立危险因素。     

北京地区急性呼吸道感染儿童中人鼻病毒感染状况研究

为了解北京地区急性呼吸道感染儿童中人鼻病毒(HRV)-A、B、C型(尤其是HRV-C)的感染状况及分子生物学特征,本研究收集了2011年1月至12月急性呼吸道感染患儿呼吸道标本703例,采用半巢式PCR方法同时进行HRV-A、B、C检测,并对HRV阳性标本进行VP4/VP2衣壳蛋白编码区基因序列测定及进化树分析。结果显示703例标本中共有54例HRV检测阳性,总阳性检出率为7.7%(54/703);HRV感染的儿童中年龄在5岁以下的占94.4%(51/54);HRV检出率在毛细支气管炎患儿中最高(23.1%),其次是哮喘(20.0%)、肺炎(11.0%)、急性支气管炎(4.3%)、急性上呼吸道感染(4.1%)患儿;序列分析表明54例HRV检测阳性标本中25例为HRV-A(46.3%,25/54),8例为HRV-B(14.8%,8/54),21例为HRV-C(38.9%,21/54);HRV同一基因型内各核苷酸序列同源性为70.0%～100.0%,不同基因型间核苷酸序列同源性为55.5%～65.8%。上述结果提示,HRV是引起北京地区儿童急性呼吸道感染(尤其是下呼吸道感染)重要的病毒病原之一;HRV-C阳性检出率与HRV-A相近,较HRV-B高;HRV各个基因型间具有较高变异性。     

重症急性呼吸道感染患儿中人鼻病毒分型检测及流行病学分析

目的:调查重症急性呼吸道感染患儿中人鼻病毒(HRV)的感染情况,初步了解不同型别HRV流行病学和临床特点。方法:收集2008年5月至2010年3月北京儿童医院重症急性呼吸道感染住院患儿鼻咽抽吸物样本259份,采用巢式PCR法,先用鼻病毒5′UTR引物筛查样本中该病毒总的感染情况,再用针对VP4-VP2区域的引物对阳性样本进行分型检测;同时对阳性样本进行其他常见呼吸道病毒共感染检测与流行病学及临床特点分析。结果:259例样本中,5′UTR初筛HRV阳性89例(34.4%),与其他常见呼吸道病毒存在共感染54例(60.7%);可通过VP4-VP2分型测序确定39例HRV-A(48.1%)、4例HRV-B(4.9%)、38例HRV-C(46.9%);HRV-A感染率最高的季节分布是秋季;HRV-A和HRV-C型感染者在临床特点上无明显差别。结论:HRV是儿童重症急性呼吸道感染的重要病原之一,新发现的HRV-C基因型感染率与HRV-A相似,但HRV在儿童重症急性呼吸道感染中的病原学意义还须进一步确认。     

我国河南与北京腹泻患儿中的星状病毒感染

人星状病毒（ＨＡｓｔＶ）是ＲＮＡ病毒的新家族,从我国河南和北京两地区的急性腹泻患儿便样中鉴定了７株星状病毒,经电镜观察病毒颗粒直径约为２８ｎｍ,表面呈五角或六角星状形态,用ＲＴ－ＰＣＲ检定为阳性,比较ＰＣＲ扩增产物的核苷酸序列构建的序列同源性树表明,１９９０年在河南流行的星状病毒主要为ＨＡｓｔＶ－１,另有一株ＨＡｓｔＶ－５,１９９６年北京流行株为ＨＡｓｔＶ－５。对ＨＡｓｔＶ患儿的临床症状进行了讨论     

布拉酵母菌散联合蒙脱石散治疗儿童迁延性腹泻的疗效观察

目的探讨布拉酵母菌散联合蒙脱石散治疗儿童迁延性腹泻的疗效观察。方法选取迁延性腹泻患儿84例,随机分为观察组和对照组。两组患儿均予以指导喂养、调整饮食,预防或纠正脱水和酸中毒等基础治疗。观察组在此基础上予以布拉酵母菌散联合蒙脱石散治疗,对照组在此基础上予以单用蒙脱石散治疗。观察两组患儿治疗后主要临床症状及体征恢复正常的时间,并比较其治疗5d后的临床疗效。结果观察组患儿的腹泻、呕吐、退热及大便常规等恢复正常时间明显短于对照组（均P〈0．05）。治疗5d后,观察组患儿的临床总有效率（95．24％）明显高于对照组（80．95％）（X^2=4．09,P〈0．05）。结论布拉酵母菌散联合蒙脱石散治疗儿童迁延性腹泻的疗效确切,能明显改善患者的临床症状及体征,缩短治疗的病程。     

Genetic characterization of Human astrovirus infection in Wuhan, People's Republic of China, 2007-2008

Human astrovirus (HAstV) was an important cause of viral gastroenteritis in infants in Wuhan city based on our previous study. The aim of the study was to investigate the nature of HAstV infection in Wuhan, People's Republic of China, especially in adults. Stool specimens were collected from 361 children and 301 adults with diarrhea from July 2007 to June 2008 and were tested for HAstV RNA by RT-PCR. The 348-bp PCR product of positive samples was further sequenced and analyzed for multiple sequence alignment and phylogenetic tree. HAstV RNA was detected in 2.33% (7/301) adults, which was significantly lower than that in children (13.57%, 49/361). HAstV-positive patients were either older than 50 years of age or younger than 3. Genetic analysis showed that the HAstV strain in adults was the same as that in children in 2007-2008. Contrarily, HAstV strains prevalent in 2007-2008 showed genetic characteristics different from those in 2004-2005 and belonged to two new groups of HAstV-1b. Thus, our data characterized HAstV infection in Wuhan 2007-2008, suggesting that HAstV infection also played an important role in adults in Wuhan, especial in patients of >50 years, and should be included for routine diagnosis in the population with diarrheal illness.     

学龄前儿童细菌性腹泻的病原学特征及PCT、CD64和sTREM-1测定的临床意义

目的:分析学龄前儿童细菌性腹泻的病原学特征及血清中降钙素原(PCT)、中性粒细胞分化群64(CD64)和髓样细胞触发受体-1(s TREM-1)检测的临床意义。方法:选择2017年1月～2018年3月南方医科大学附属东莞市人民医院收治的214例学龄前感染性腹泻患儿为研究对象,包括细菌性腹泻(细菌组,n=101)和病毒性腹泻(病毒组,n=113),以同期入院行儿童保健的113例健康学龄前儿童为对照组。统计细菌组患儿的病原菌种类,比较两组腹泻患儿与对照组儿童血清PCT、CD64和s TREM-1水平的差异、三种血清学指标的阳性率及三种指标对细菌性腹泻患儿的诊断效能。结果:101例细菌性腹泻患儿粪便培养的细菌菌株中,共培养菌株213株,大肠埃希菌、志贺菌属和沙门菌属阳性率最高,分别占41.78%、23.01%和15.96%;细菌组的血清PCT、CD64和s TREM-1水平及阳性率均高于病毒组和对照组,且病毒组的s TREM-1水平高于对照组(P0.05);三种指标的灵敏度、特异度、阳性预测值和阴性预测值比较,差异无统计学意义(P0.05)。结论:学龄前儿童细菌性腹泻的致病菌主要为革兰阴性菌,PCT、CD64和s TREM-1均可作为判断细菌性腹泻的早期敏感指标。     

33例普通型新型冠状病毒肺炎的临床特征分析

摘要 目的：总结33例普通型新型冠状病毒肺炎(coronavirus disease 2019,COVID-19)患者的临床特征,为临床治疗提供指导。方法：回顾性分析我院收治的33例COVID-19患者的临床资料,对其特征进行分析。结果：31例患者有流行病学接触史,其中16例有家庭聚集情况。最常见的临床症状为发热、咳嗽等上呼吸道感染症状,少见可出现腹泻、上腹部不适,罕见头痛、视神经脊髓炎。患者治疗后实验室大部分指标均改善明显,但淋巴细胞以及心肌损伤标记物高敏心肌肌钙蛋白T(high sensitive cardiac troponin,hs-cTnT)改善稍晚。97.0 %的患者胸部CT中以磨玻璃影为主,但这与普通病毒性肺炎无明显区别,老年患者可合并胸腔积液。结论：在西安地区收治的COVID-19患者多数有明确的流行病学史,临床表现主要为发热咳嗽,但也可累及消化系统及神经系统出现症状。患者经治疗后实验室指标大部分改善明显,肺部影像学以磨玻璃样改变居多,也可出现胸腔积液。普通型患者经治疗后预后良好。     

Dengue infection in children in Ratchaburi, Thailand: a cohort study. II. Clinical manifestations

Background

Dengue infection is one of the most important mosquito-borne diseases. More data regarding the disease burden and the prevalence of each clinical spectrum among symptomatic infections and the clinical manifestations are needed. This study aims to describe the incidence and clinical manifestations of symptomatic dengue infection in Thai children during 2006 through 2008.

Study Design

This study is a school-based prospective open cohort study with a 9,448 person-year follow-up in children aged 3–14 years. Active surveillance for febrile illnesses was done in the studied subjects. Subjects who had febrile illness were asked to visit the study hospital for clinical and laboratory evaluation, treatment, and serological tests for dengue infection. The clinical data from medical records, diary cards, and data collection forms were collected and analyzed.

Results

Dengue infections were the causes of 12.1% of febrile illnesses attending the hospital, including undifferentiated fever (UF) (49.8%), dengue fever (DF) (39.3%) and dengue hemorrhagic fever (DHF) (10.9%). Headache, anorexia, nausea/vomiting and myalgia were common symptoms occurring in more than half of the patients. The more severe dengue spectrum (i.e., DHF) had higher temperature, higher prevalence of nausea/vomiting, abdominal pain, rash, diarrhea, petechiae, hepatomegaly and lower platelet count. DHF cases also had significantly higher prevalence of anorexia, nausea/vomiting and abdominal pain during day 3–6 and diarrhea during day 4–6 of illness. The absence of nausea/vomiting, abdominal pain, diarrhea, petechiae, hepatomegaly and positive tourniquet test may predict non-DHF.

Conclusion

Among symptomatic dengue infection, UF is most common followed by DF and DHF. Some clinical manifestations may be useful to predict the more severe disease (i.e., DHF). This study presents additional information in the clinical spectra of symptomatic dengue infection.     

Feline panleukopenia viral infection in cats: Application of some molecular methods used for its diagnosis

Forty diseased cats and seven healthy control cats from different sex, ages and breeds had examined clinically to confirm presence or absence of clinical symptoms of Feline panleukopenia disease (FP). Several tools including ELISA, gene expression analysis (qRT-PCR), DNA fragmentation test and apoptosis assay were conducted to determine the Feline panleukopenia disease in cat tissues. Clinical symptoms in the form of depression, fever, anorexia, vomiting, diarrhea, dehydration, anaemia and leucopenia were recorded in the diseased cats while no clinical sings were observed in control healthy cats. ELISA results showed that all of diseased (n?=?40) cats were positive while control cats (n?=?7) were negative for FP viral antigen. After carrying out of ELISA assay, supportive treatment trials including fluid therapy, immunostimulant, antibiotics to overcome dehydration, restoring electrolytes imbalances, combating secondary bacterial infection were conducted but all diseased cats were died and control cats exposed to soft death. Gene expression analysis detected high levels of FP viral gene in several cat tissues in which ilium exhibited high viral expression levels compared with jejunum. Also, viral expression levels in jejunum were higher than in mesenteric lymph nodes. In addition, viral expression levels were not detected in tissues of control cats. The results of the DNA fragmentation assay observed that DNA extracted from different tissues of infected cats exhibited damaged DNA bands as compared with DNA of control cats. DNA fragmentation rates in infected tissues increased significantly (P?<?0.01), the highest rates were showed in ilium and jejunum tissue than in mesenteric lymph nodes. Determination of apoptosis in cat tissues showed that rate of apoptosis/necrosis increased significantly (P?<?0.05) in infected cats tissues in comparison to control cats. Moreover the highest apoptotic ratios of infected cats were observed in ilium and jejunum tissues compared with mesenteric lymph nodes.     

Type I Interferon Response Is Delayed in Human Astrovirus Infections

Type I interferon (IFN) activation and its subsequent effects are important in the response to viral infections. Here we show that human astroviruses (HAstVs), which are important agents of acute gastroenteritis in children, induce a mild and delayed IFN response upon infecting CaCo-2 cells. Although IFN-β mRNA is detected within infected cells and supernatant from infected cells show antiviral activity against the replication of other well-known IFN-sensitive viruses, these responses occur at late stages of infection once genome replication has taken place. On the other hand, HAstV replication can be partially reduced by the addition of exogenous IFN, and inhibition of IFN activation by BX795 enhances viral replication, indicating that HAstVs are IFN-sensitive viruses. Finally, different levels of IFN response were observed in cells infected with different HAstV mutants with changes in the hypervariable region of nsP1a/4, suggesting that nsP1a/4 genotype may potentially have clinical implications due to its correlation with the viral replication phenotype and the antiviral responses induced within infected cells.